LOCUS       sequence-389540-        6282 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian inducible expression vector for myc-tagged SPOP.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6282)
  AUTHORS   Sabri N, Cuneo MJ, Marzahn MR, Lee J, Bouchard JJ, Gullulu O, 
            Vaithiyalingam S, Borgia MB, Schmit J, Mittag T
  TITLE     Reduction of oligomer size modulates the competition between cluster
            formation and phase separation of the tumor suppressor SPOP.
  JOURNAL   J Biol Chem. 2023 Dec;299(12):105427. doi: 
            10.1016/j.jbc.2023.105427. Epub 2023 Nov 4.
  PUBMED    37926283
REFERENCE   2  (bases 1 to 6282)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "J Biol 
            Chem."; date: "2023-12"; volume: "299(12):105427. doi"; pages: " 
            10.1016/j.jbc.2023.105427. Epub 2023 Nov 4"
COMMENT     Sequence Label: pCDNA4/TO-SPOP-myc (BACK)
FEATURES             Location/Qualifiers
     source          1..6282
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(44..63)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        235..614
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        615..818
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     769..789
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     protein_bind    820..838
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    841..859
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     primer_bind     869..893
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     CDS             2205..2234
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             2250..2267
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(2290..2307)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2296..2520
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      2566..2994
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2653..2672)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2863..2884
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3003..3023)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3008..3337
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3188..3323
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     3250..3269
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     promoter        3385..3432
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             3451..3825
                     /codon_start=1
                     /gene="Sh ble from Streptoalloteichus hindustanus"
                     /product="antibiotic-binding protein"
                     /label=BleoR
                     /note="confers resistance to bleomycin, phleomycin, and 
                     Zeocin(TM)"
                     /translation="MAKLTSAVPVLTARDVAGAVEFWTDRLGFSRDFVEDDFAGVVRDD
                     VTLFISAVQDQVVPDNTLAWVWVRGLDELYAEWSEVVSTNFRDASGPAMTEIGEQPWGR
                     EFALRDPAGNCVHFVAEEQD"
     polyA_signal    3955..4076
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3992..4011)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4046..4065
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4125..4141)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4125..4141)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4138..4160)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4149..4165
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     promoter        complement(join(4173..4179,4180..4197,4198..4203))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4218..4239
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4356..4373)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4527..5115)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4607..4626)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5286..6077,6078..6146))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     5909..5928
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6147..6251)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gacggatcgg gagatctccc gatcccctat ggtgcactct cagtacaatc tgctctgatg
       61 ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg
      121 cgagcaaaat ttaagctaca acaaggcaag gcttgaccga caattgcatg aagaatctgc
      181 ttagggttag gcgttttgcg ctgcttcgcg atgtacgggc cagatatacg cgttgacatt
      241 gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata
      301 tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc
      361 cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc
      421 attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt
      481 atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt
      541 atgcccagta catgacctta tgggactttc ctacttggca gtacatctac gtattagtca
      601 tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg
      661 actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc
      721 aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg
      781 gtaggcgtgt acggtgggag gtctatataa gcagagctct ccctatcagt gatagagatc
      841 tccctatcag tgatagagat cgtcgacgag ctcgtttagt gaaccgtcag atcgcctgga
      901 gacgccatcc acgctgtttt gacctccata gaagacaccg ggaccgatcc agcctccgga
      961 ctctagcgtt taaacttaag cttggtaccg agctcggatc cactagtcca gtgtggtgga
     1021 attctgcaga tgtgcaatcc tgcagatatc caatgtcaag ggttccaagt cctccacctc
     1081 cggcagaaat gtcgagtggt cccgttgccg agagctggtg ctatacacag atcaaggtag
     1141 tgaagttctc ctacatgtgg accatcaata actttagttt ttgccgggag gaaatgggtg
     1201 aagtcataaa aagttcaact ttttcatcag gagccaatga taaactgaaa tggtgtttgc
     1261 gagtaaaccc gaaagggcta gatgaagaaa gcaaagatta cctgtcgctt tacctgttgt
     1321 tggtcagctg tccaaagagt gaagttcggg caaagttcaa gttctccatc ctgaatgcca
     1381 agggagaaga aaccaaagct atggagagtc agcgagctta taggttcgtg caaggcaaag
     1441 actggggatt caaaaaattc atccgcagag attttctctt ggacgaggcc aatgggcttc
     1501 tccctgatga caagctcacc ctcttctgtg aggtgagtgt agtacaagac tctgtcaata
     1561 tttctgggca gaatactatg aacatggtaa aggttcctga atgccggttg gcagatgagt
     1621 taggagggct gtgggagaac tctcgattca ccgactgctg cttgtgtgtg gctggtcaag
     1681 aattccaggc tcacaaggct atcttagcag ctcggtctcc agtcttcagt gccatgtttg
     1741 aacatgaaat ggaggagagc aaaaagaatc gggttgagat taacgacgtg gagcctgagg
     1801 tgtttaagga gatgatgtgc ttcatctaca cggggaaggc gccgaacctc gacaagatgg
     1861 ctgacgactt gctggcagct gccgataagt acgccctgga gcgtctgaag gtcatgtgtg
     1921 aggacgccct ctgcagtaac ctctctgtgg agaacgctgc agagattctc atcctggctg
     1981 acctccacag cgcagatcag ctgaagacgc aggcagttga tttcatcaac tatcatgctt
     2041 cggatgtctt ggagacctct gggtggaagt caatggtggt gtcacatccc cacttagtgg
     2101 cagaagcaga gcgttctctg gcttcagcac agtgcccttt cctggggccc ccacgcaaac
     2161 gcctgaagca atcctggcgg ccgctcgagt ctagagggcc cttcgaacaa aaactcatct
     2221 cagaagagga tctgaatatg cataccggtc atcatcacca tcaccattga gtttaaaccc
     2281 gctgatcagc ctcgactgtg ccttctagtt gccagccatc tgttgtttgc ccctcccccg
     2341 tgccttcctt gaccctggaa ggtgccactc ccactgtcct ttcctaataa aatgaggaaa
     2401 ttgcatcgca ttgtctgagt aggtgtcatt ctattctggg gggtggggtg gggcaggaca
     2461 gcaaggggga ggattgggaa gacaatagca ggcatgctgg ggatgcggtg ggctctatgg
     2521 cttctgaggc ggaaagaacc agctggggct ctagggggta tccccacgcg ccctgtagcg
     2581 gcgcattaag cgcggcgggt gtggtggtta cgcgcagcgt gaccgctaca cttgccagcg
     2641 ccctagcgcc cgctcctttc gctttcttcc cttcctttct cgccacgttc gccggctttc
     2701 cccgtcaagc tctaaatcgg gggctccctt tagggttccg atttagtgct ttacggcacc
     2761 tcgaccccaa aaaacttgat tagggtgatg gttcacgtag tgggccatcg ccctgataga
     2821 cggtttttcg ccctttgacg ttggagtcca cgttctttaa tagtggactc ttgttccaaa
     2881 ctggaacaac actcaaccct atctcggtct attcttttga tttataaggg attttgccga
     2941 tttcggccta ttggttaaaa aatgagctga tttaacaaaa atttaacgcg aattaattct
     3001 gtggaatgtg tgtcagttag ggtgtggaaa gtccccaggc tccccagcag gcagaagtat
     3061 gcaaagcatg catctcaatt agtcagcaac caggtgtgga aagtccccag gctccccagc
     3121 aggcagaagt atgcaaagca tgcatctcaa ttagtcagca accatagtcc cgcccctaac
     3181 tccgcccatc ccgcccctaa ctccgcccag ttccgcccat tctccgcccc atggctgact
     3241 aatttttttt atttatgcag aggccgaggc cgcctctgcc tctgagctat tccagaagta
     3301 gtgaggaggc ttttttggag gcctaggctt ttgcaaaaag ctcccgggag cttgtatatc
     3361 cattttcgga tctgatcagc acgtgttgac aattaatcat cggcatagta tatcggcata
     3421 gtataatacg acaaggtgag gaactaaacc atggccaagt tgaccagtgc cgttccggtg
     3481 ctcaccgcgc gcgacgtcgc cggagcggtc gagttctgga ccgaccggct cgggttctcc
     3541 cgggacttcg tggaggacga cttcgccggt gtggtccggg acgacgtgac cctgttcatc
     3601 agcgcggtcc aggaccaggt ggtgccggac aacaccctgg cctgggtgtg ggtgcgcggc
     3661 ctggacgagc tgtacgccga gtggtcggag gtcgtgtcca cgaacttccg ggacgcctcc
     3721 gggccggcca tgaccgagat cggcgagcag ccgtgggggc gggagttcgc cctgcgcgac
     3781 ccggccggca actgcgtgca cttcgtggcc gaggagcagg actgacacgt gctacgagat
     3841 ttcgattcca ccgccgcctt ctatgaaagg ttgggcttcg gaatcgtttt ccgggacgcc
     3901 ggctggatga tcctccagcg cggggatctc atgctggagt tcttcgccca ccccaacttg
     3961 tttattgcag cttataatgg ttacaaataa agcaatagca tcacaaattt cacaaataaa
     4021 gcattttttt cactgcattc tagttgtggt ttgtccaaac tcatcaatgt atcttatcat
     4081 gtctgtatac cgtcgacctc tagctagagc ttggcgtaat catggtcata gctgtttcct
     4141 gtgtgaaatt gttatccgct cacaattcca cacaacatac gagccggaag cataaagtgt
     4201 aaagcctggg gtgcctaatg agtgagctaa ctcacattaa ttgcgttgcg ctcactgccc
     4261 gctttccagt cgggaaacct gtcgtgccag ctgcattaat gaatcggcca acgcgcgggg
     4321 agaggcggtt tgcgtattgg gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg
     4381 gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg cggtaatacg gttatccaca
     4441 gaatcagggg ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac
     4501 cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc gcccccctga cgagcatcac
     4561 aaaaatcgac gctcaagtca gaggtggcga aacccgacag gactataaag ataccaggcg
     4621 tttccccctg gaagctccct cgtgcgctct cctgttccga ccctgccgct taccggatac
     4681 ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc atagctcacg ctgtaggtat
     4741 ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag
     4801 cccgaccgct gcgccttatc cggtaactat cgtcttgagt ccaacccggt aagacacgac
     4861 ttatcgccac tggcagcagc cactggtaac aggattagca gagcgaggta tgtaggcggt
     4921 gctacagagt tcttgaagtg gtggcctaac tacggctaca ctagaagaac agtatttggt
     4981 atctgcgctc tgctgaagcc agttaccttc ggaaaaagag ttggtagctc ttgatccggc
     5041 aaacaaacca ccgctggtag cggtggtttt tttgtttgca agcagcagat tacgcgcaga
     5101 aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc tcagtggaac
     5161 gaaaactcac gttaagggat tttggtcatg agattatcaa aaaggatctt cacctagatc
     5221 cttttaaatt aaaaatgaag ttttaaatca atctaaagta tatatgagta aacttggtct
     5281 gacagttacc aatgcttaat cagtgaggca cctatctcag cgatctgtct atttcgttca
     5341 tccatagttg cctgactccc cgtcgtgtag ataactacga tacgggaggg cttaccatct
     5401 ggccccagtg ctgcaatgat accgcgagac ccacgctcac cggctccaga tttatcagca
     5461 ataaaccagc cagccggaag ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc
     5521 atccagtcta ttaattgttg ccgggaagct agagtaagta gttcgccagt taatagtttg
     5581 cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct
     5641 tcattcagct ccggttccca acgatcaagg cgagttacat gatcccccat gttgtgcaaa
     5701 aaagcggtta gctccttcgg tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta
     5761 tcactcatgg ttatggcagc actgcataat tctcttactg tcatgccatc cgtaagatgc
     5821 ttttctgtga ctggtgagta ctcaaccaag tcattctgag aatagtgtat gcggcgaccg
     5881 agttgctctt gcccggcgtc aatacgggat aataccgcgc cacatagcag aactttaaaa
     5941 gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct caaggatctt accgctgttg
     6001 agatccagtt cgatgtaacc cactcgtgca cccaactgat cttcagcatc ttttactttc
     6061 accagcgttt ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg
     6121 gcgacacgga aatgttgaat actcatactc ttcctttttc aatattattg aagcatttat
     6181 cagggttatt gtctcatgag cggatacata tttgaatgta tttagaaaaa taaacaaata
     6241 ggggttccgc gcacatttcc ccgaaaagtg ccacctgacg tc
//
